番茄红素抑制铁死亡改善APAP诱导鸡LMH肝细胞损伤的作用及其机制  

The effect and mechanism of lycopene on improving APAP induced chicken LMH liver cell injury by inhibiting ferroptosis

作　　者：王坤丽 闫一博 邱艺康 关医博 聂少琪 姜雨辰 李梓怡 杜宇[1] 张磊[1] 康宇 WANG Kunli;YAN Yibo;QIU Yikang;GUAN Yibo;NIE Shaoqi;JIANG Yuchen;LI Ziyi;DU Yu;ZHANG Lei;KANG Yu(College of Veterinary Medicine,Henan University of Animal Husbandry and Economy,Zhengzhou Henan 450046)

机构地区：[1]河南牧业经济学院动物医药学院,河南郑州450046

出　　处：《现代牧业》2024年第4期31-37,共7页Modern Animal Husbandry

基　　金：河南牧业经济学院博士启动基金项目(M4030067)。

摘　　要：基于铁死亡信号通路探讨番茄红素(Lyc)对对乙酰氨基酚(APAP)诱导的鸡LMH肝细胞的保护作用及潜在机制。以APAP诱导鸡LMH肝细胞损伤为模型,采用细胞计数法(CCK-8)测定不同剂量Lyc预处理后LMH肝细胞存活率;通过生化试剂盒检测细胞培养液上清中丙氨酸氨基转移酶(ALT)和天门冬氨酸氨基转移酶(AST)的活性;使用FerroOrange探针检测细胞中Fe2+水平;Liperfluo探针检测脂质过氧化水平;流式细胞术检测细胞中活性氧(ROS)水平;比色法检测培养液上清中乳酸脱氢酶(LDH)水平和细胞中谷胱甘肽(GSH)水平;免疫印迹测定长链脂酰辅酶A合成酶4(ACSL4)和谷胱甘肽过氧化酶4(GPX4)蛋白表达;免疫荧光检测沉默信息调节因子-1(Sirt1)和核因子E2相关因子2(Nrf2)表达。与对照组比较,APAP组鸡LMH肝细胞活力明显降低(P<0.05),细胞培养液上清中ALT和AST活性升高(P<0.05);Lyc干预后鸡LMH肝细胞活力明显升高,ALT和AST活性降低,与APAP组相比有显著差异(P<0.05);与对照组比较,APAP组细胞中Fe2+、ROS、脂质过氧化和LDH水平以及ACSL4蛋白表达均升高(P<0.05);此外,APAP组鸡LMH肝细胞中GSH水平以及Sirt1、Nrf2和GPX4蛋白表达降低,与对照组比较有显著差异(P<0.05);与APAP组相比,Lyc组鸡LMH肝细胞中Fe2+、ROS和脂质过氧化水平、LDH水平和ACSL4蛋白表达降低,GSH水平以及Sirt1、Nrf2和GPX4蛋白表达增加,均有显著性差异(P<0.05)。Sirt1抑制剂EX527减弱了Lyc对APAP处理条件下鸡LMH肝细胞铁死亡的抑制作用。Lyc能缓解APAP诱导的鸡LMH肝细胞铁死亡,其可能与调控Sirt1/Nrf2信号有关。This study explores the protective effect of lycopene(Lyc)on acetaminophen(APAP)-induced chicken LMH liver cells and its potential mechanism based on the ferroptosis signaling pathway.Using APAP-induced chicken LMH liver cell damage as a model,the cell counting method(CCK-8)was used to determine the survival rate of chicken LMH liver cells after different doses of Lyc pretreatment.Detect the activity of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in the supernatant of cell culture medium using the biochemical assay kits.FerroOrange probe was used to detect Fe2+levels in cells.Liperfluo probe was used to detect lipid peroxidation levels.Flow cytometry was used to detect reactive oxygen species(ROS)levels in cells.The colorimetric method was used to detect lactate dehydrogenase(LDH)levels and glutathione(GSH)levels in culture supernatant.Immunoblotting was used to measure the protein expression of long-chain acyl CoA synthase 4(ACSL4)and glutathione peroxidase 4(GPX4),and immunofluorescence was used to detect the expression of silencing information regulatory factor-1(Sirt1)and nuclear factor E2 related factor 2(Nrf2).Compared with the control group,the APAP group showed a significant decrease in chicken LMH liver cell viability(P<0.05),and an increase in ALT and AST activities in the cell culture supernatant(P<0.05).After Lyc intervention,chicken LMH liver cell viability significantly increased,while ALT and AST activities decreased,with significant differences compared to the APAP group(P<0.05).Compared with the control group,the levels of Fe2+,ROS,lipid peroxidation,LDH,and ACSL4 protein expression in APAP group cells were increased(P<0.05).In addition,the levels of GSH and the expression of Sirt1,Nrf2,and GPX4 proteins in chicken LMH liver cells in the APAP group were significantly reduced compared to the control group(P<0.05).Compared with the APAP group,the levels of Fe2+,ROS,and lipid peroxidation in chicken LMH liver cells,as well as the levels of LDH and ACSL4 protein expression,decreas

关 键 词：番茄红素 对乙酰氨基酚 铁死亡 肝细胞 Sirt1/Nrf2 

分 类 号：S816[农业科学—饲料科学]